Heat degradation of eukaryotic and bacterial DNA: an experimental model for paleomicrobiology.

ABSTRACT:

Background

Theoretical models suggest that DNA degradation would sharply limit the PCR-based detection of both eukaryotic and prokaryotic DNA within ancient specimens. However, the relative extent of decay of eukaryote and prokaryote DNA over time is a matter of debate. In this study, the murine macrophage cell line J774, alone or infected with Mycobacterium smegmatis bacteria, were killed after exposure to 90°C dry heat for intervals ranging from 1 to 48 h in order to compare eukaryotic cells, extracellular bacteria and intracellular bacteria. The sizes of the resulting mycobacterial rpoB and murine rpb2 homologous gene fragments were then determined by real-time PCR and fluorescent probing.

Findings

The cycle threshold (Ct) values of PCR-amplified DNA fragments from J774 c

SUBMITTER: Nguyen-Hieu T 

PROVIDER: S-EPMC3532149 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature